This research will encompass studies of the enzymology of mammalian spermatozoan acrosomes with special emphasis on the enzyme systems that are involved in fertilization. These studies will use zymographic and histochemical techniques to characterize and thereby identify certain enzymes of bull, ram and rabbit acrosomes after electrophoresis of isolated acrosomal materials on polyacrylamide gels to separate the enzymes from in situ enzyme inhibitors. The specific biochemical characteristics of each acrosomal enzyme such as zymographic staining, carbohydrate of lipid components, and electrophoretic mobility will be used to determine the effectiveness of isolation attempts using column chromatography, affinity chromatography, isoelectric focusing, and preparative-electrophoresis. The biochemical characteristics of the enzymes will be used to examine the relationships between the enzymes and certain physiological processes such as spermatozoan maturation, capacitation, and senescence. The effect of certain hormones, parahormones, and other substances on the macromolecular components of spermatozoan acrosomes will be determined. The antigenic properties of purified acrosomal proteins will be used for immunological localization of the proteins in spermatozoa. The immunological effect of purified acrosomal proteins on spermatozoan fertility will be examined also. Purpose of these studies is to obtain fundamental biochemical and physiological information on mammalian acrosomal enzyme systems thereby extending the possibilities for development of new contraceptive methods.